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β防御素作为生物标志物:在 LPS 刺激的马外周血单个核细胞和正常、无菌和可能感染性马关节液中可检测到。

Beta defensins as biomarkers: detectable in LPS-stimulated equine peripheral blood mononuclear cells and normal, aseptic, and probable septic equine synovial fluid.

机构信息

Department of Pathobiology and Diagnostic Investigation, College of Veterinary Medicine, Michigan State University, East Lansing, MI.

Department of, Large Animal Clinical Sciences, College of Veterinary Medicine, Michigan State University, East Lansing, MI.

出版信息

Am J Vet Res. 2022 Jul 20;83(9):ajvr.21.12.0204. doi: 10.2460/ajvr.21.12.0204.

Abstract

OBJECTIVES

Diagnosis of equine septic arthritis is not straightforward, and increasing time between onset, diagnosis, and treatment can have serious consequences for quality of life. Defensins are used in diagnosis of human joint infection. The presence of beta defensins (BDs) in equine synovial fluid and their utility as a biomarker of sepsis has not been investigated; therefore, our objectives were to (1) compare in vitro gene expression of lipopolysaccharide (LPS)-stimulated equine neutrophils to unstimulated neutrophils and (2) compare BD protein expression from normal, aseptically inflamed, and septic equine joints.

ANIMALS

5 horses for isolated neutrophil BD expression and 21 synovial fluid samples from 14 horses.

PROCEDURES

RT-qPCR analysis was performed for BD gene expression of stimulated and unstimulated equine peripheral neutrophils. BD protein expression was evaluated from equine joints with no disease, aseptic inflammation, and septic inflammation using a commercial ELISA designed for horses and analyzed with a Kruskal-Wallis test (significant at P < .05).

RESULTS

A significant increase was noted in expression of BD-3 in LPS stimulated as compared to unstimulated neutrophils. There were no significant differences in BD expression noted between joints with no disease, aseptic inflammation, and septic inflammation. Low case numbers and different types of cases in the aseptic inflammation group were main limitations. BD expression patterns in samples from stimulated equine peripheral neutrophils and synovial fluid were identified.

CLINICAL RELEVANCE

BDs are detectable in equine synovial fluid and can be stimulated from peripheral neutrophils. Further examination is needed to define their role as biomarkers of joint disease.

摘要

目的

马属动物脓毒性关节炎的诊断并不简单,发病、诊断和治疗之间的时间延长可能会对生活质量产生严重影响。防御素用于人类关节感染的诊断。马关节滑液中β防御素(BDs)的存在及其作为脓毒症生物标志物的用途尚未得到研究;因此,我们的目的是(1)比较脂多糖(LPS)刺激的马中性粒细胞与未刺激的中性粒细胞的体外基因表达,(2)比较正常、无菌性炎症和脓毒性马关节的 BD 蛋白表达。

动物

5 匹用于分离中性粒细胞 BD 表达的马和 14 匹马的 21 个关节滑液样本。

程序

对 LPS 刺激和未刺激的马外周中性粒细胞的 BD 基因表达进行 RT-qPCR 分析。使用专为马设计的商业 ELISA 评估无疾病、无菌性炎症和脓毒性炎症的马关节中的 BD 蛋白表达,并使用 Kruskal-Wallis 检验进行分析(P<.05 有统计学意义)。

结果

与未刺激的中性粒细胞相比,LPS 刺激的 BD-3 表达显著增加。无疾病、无菌性炎症和脓毒性炎症关节之间的 BD 表达无显著差异。无菌性炎症组病例数量少且病例类型不同是主要限制因素。确定了从刺激的马外周中性粒细胞和关节滑液样本中鉴定出的 BD 表达模式。

临床相关性

BDs 可在马关节滑液中检测到,并可从外周中性粒细胞中刺激产生。需要进一步研究来确定它们作为关节疾病生物标志物的作用。

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